MONITORING OF PROTEIN CONFORMATION BY HIGH-PERFORMANCE SIZE-EXCLUSION LIQUID-CHROMATOGRAPHY AND SCANNING DIODE-ARRAY 2ND-DERIVATIVE UV ABSORPTION-SPECTROSCOPY

被引：16

作者：

ACKLAND, CE ^{[1
]}

BERNDT, WG ^{[1
]}

FREZZA, JE ^{[1
]}

LANDGRAF, BE ^{[1
]}

PRITCHARD, KW ^{[1
]}

CIARDELLI, TL ^{[1
]}

机构：

[1] DARTMOUTH COLL,HITCHCOCK MED CTR,DARTMOUTH MED SCH,DEPT PHARMACOL,HANOVER,NH 03756

来源：

JOURNAL OF CHROMATOGRAPHY | 1991年 / 540卷 / 1-2期

关键词：

D O I：

10.1016/S0021-9673(01)88808-7

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

Genetic methods now allow the rapid production of mutant proteins for structure-function analysis. To properly interpret any change in biologic activity resulting from modification in primary sequence, it is essential to monitor conformational changes resulting from mutations. Several methods allow low-resolution protein conformational analysis. One method, second-derivative UV absorption spectroscopy, is particularly useful for proteins containing tyrosine and/or tryptophan residues. Using high-performance size-exclusion liquid chromatography and scanning diode array detection we have demonstrated that it is possible to monitor the degree of aggregation as well as conformational perturbation for a series of interleukin-2 structural mutants. Furthermore, the combination of high-performance liquid chromatography and second-derivative UV absorption spectroscopy avoids a potential artifactual contribution in non-chromatographic analysis due to protein aggregation.

引用

页码：187 / 198

页数：12

共 20 条

[1] HELIX GEOMETRY IN PROTEINS [J].

BARLOW, DJ ;

THORNTON, JM .

JOURNAL OF MOLECULAR BIOLOGY, 1988, 201 (03) :601-619

[2] STABILIZATION OF AN ASSOCIATED FOLDING INTERMEDIATE OF BOVINE GROWTH-HORMONE BY SITE-DIRECTED MUTAGENESIS [J].

BREMS, DN ;

PLAISTED, SM ;

HAVEL, HA ;

TOMICH, CSC .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1988, 85 (10) :3367-3371

[3] STRUCTURE-ACTIVITY STUDIES OF INTERLEUKIN-2 [J].

COHEN, FE ;

KOSEN, PA ;

KUNTZ, ID ;

EPSTEIN, LB ;

CIARDELLI, TL ;

SMITH, KA .

SCIENCE, 1986, 234 (4774) :349-352

[4] INTERLEUKIN-2 SELF-ASSOCIATION [J].

FLEISCHMANN, JD ;

WENTWORTH, D ;

VALENCIC, F ;

IMBEMBO, AL ;

KOEHLER, KA .

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1988, 152 (02) :879-885

[5] USE OF SCANNING DIODE-ARRAY DETECTOR WITH REVERSED-PHASE MICROBORE COLUMNS FOR THE REAL-TIME SPECTRAL-ANALYSIS OF AROMATIC-AMINO-ACIDS IN PEPTIDES AND PROTEINS AT THE SUBMICROGRAM LEVEL - APPLICATIONS TO PEPTIDE AND PROTEIN MICROSEQUENCIN [J].

GREGO, B ;

NICE, EC ;

SIMPSON, RJ .

JOURNAL OF CHROMATOGRAPHY, 1986, 352 :359-368

[6] REVERSIBLE SELF-ASSOCIATION OF BOVINE GROWTH-HORMONE DURING EQUILIBRIUM UNFOLDING [J].

HAVEL, HA ;

KAUFFMAN, EW ;

PLAISTED, SM ;

BREMS, DN .

BIOCHEMISTRY, 1986, 25 (21) :6533-6538

[7]

ICHIKAWA T, 1981, CHEM PHARM BULL, V29, P438

[8]

KUNKEL TA, 1987, METHOD ENZYMOL, V154, P367

[9]

LANDGRAF B, 1991, IN PRESS PROTEINS ST, V9

[10]

MATTHEWS CW, 1988, METHOD ENZYMOL, V154, P498

← 1 2 →